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Volume 24, Number 7—July 2018
Research

Diversity of Influenza A(H5N1) Viruses in Infected Humans, Northern Vietnam, 2004–2010

Hirotaka Imai1, Jorge M. Dinis1, Gongxun Zhong, Louise H. Moncla, Tiago J.S. Lopes, Ryan McBride, Andrew J. Thompson, Wenjie Peng, Mai thi Q. Le, Anthony Hanson, Michael Lauck, Yuko Sakai-Tagawa, Shinya Yamada, Julie Eggenberger, David H. O’Connor, Yasuo Suzuki, Masato Hatta, James C. Paulson, Gabriele Neumann, and Yoshihiro KawaokaComments to Author 
Author affiliations: University of Wisconsin, Madison, Wisconsin, USA (H. Imai, J.M. Dinis, G. Zhong, L.H. Moncla, T.J.S. Lopes, A. Hanson, M. Lauck, J. Eggenberger, D.H. O’Connor, M. Hatta, G. Neumann, T.C. Friedrich, Y. Kawaoka); The Scripps Research Institute, La Jolla, California, USA (R. McBride, A.J. Thompson, W. Peng, J.C. Paulson); National Institute of Hygiene and Epidemiology, Hanoi, Vietnam (M.t.Q. Le); University of Tokyo, Tokyo, Japan (Y. Sakai-Tagawa, S. Yamada, Y. Kawaoka); Chubu University, Kasugai, Japan (Y. Suzuki)

Main Article

Figure 5

Effect of amino acid variations in polymerases and NP on viral polymerase activity in influenza A(H5N1) virus isolates from humans, northern Vietnam, 2004–2010. 293T cells were transfected with plasmids encoding the viral replication complex (PB2, PB1, PA, and NP), with a plasmid for the expression of an influenza virus mini-genome that encodes the firefly luciferase gene, and with a plasmid encoding Renilla luciferase (transfection control). If 2 or 3 isolate numbers are listed, we tested the m

Figure 5. Effect of amino acid variations in polymerases and NP on viral polymerase activity in influenza A(H5N1) virus isolates from humans, northern Vietnam, 2004–2010. 293T cells were transfected with plasmids encoding the viral replication complex (PB2, PB1, PA, and NP), with a plasmid for the expression of an influenza virus mini-genome that encodes the firefly luciferase gene, and with a plasmid encoding Renilla luciferase (transfection control). If 2 or 3 isolate numbers are listed, we tested the major sequence variant, which is identical among the samples. The cells were incubated at 33°C (A) or at 37°C (B) for 24 h, and firefly and Renilla luciferase activities were measured by use of the Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA). The firefly luciferase values were divided by the Renilla luciferase values to normalize for variances in transfection efficiency. The experiments (each in triplicate) were independently repeated twice. The mean relative viral polymerase activities plus SDs of each independent experiment are shown as black and white bars. The viral polymerase activity of the respective majority variant was set to 100%. NP, nucleocapsid; PA, polymerase acidic; PB, polymerase basic; *p<0.05; **p<0.01 (both by Dunnett test).

Main Article

1These authors contributed equally to this article.

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